Oligonucleotide for detection and identification of Mycobacteria

ABSTRACT

This invention relates to oligonucleotides sequence of probes or primers for detection or identication of Mycobacterium. In the claimed invention, oligonucleotide sequences of ITS (Internal Transcribing Spacer Region) from  M. fortuitium, M. chelonae, M. abscessus, M. vaccae, M. flavescence, M. Asiaticum, M. porcinum, M. acapulcensis  and  M. diernhoferi  have been identified. Using these ITS sequences, PCR primers or hybridization probes for detection or identication of Mycobacterium have been developed and presented as seq ID: 10 to seq ID: 241.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to oligonucleotides that can be used fordetection and identification of mycobacteria. More particularly, thepresent invention identifies the nucleotide sequence of ITS (InternalTranscribed Spacer region) of non-tuberculosis mycobacteria,Mycobacterium fortuitum, Mycobacterium chelonae, Mycobacteriumabscessus, Mycobacterium vaccae, Mycobacterium flavescens, Mycobacteriumasiaticum, Mycobacterium porcinum, Mycobacterium acapulcensis andMycobacterium diernhoferi, and using the nucleotide sequences, itprovides oligonucleotide primers or probes used for detection andidentification of mycobacteria.

2. Description of the Related Art

A Even though the number of patients of tuberculosis has steadilydecreased in these days, about 8 million patients have come out andabout 3 million patients died of tuberculosis in a year. Moreover, inunderdeveloped countries, inadequate treatment and lack of drugs fortuberculosis increase chronic carriers of drug-resistant bacteria. In1980's, spread of AIDS has increased patients of tuberculosis even inadvanced countries. In this condition, it is expected that about twelvemillion patents of tuberculosis would newly come out in the year 2000(J. P. Natain, M. C. Raviglione, and A. Kochi, Tubercle and LungDisease, 73: 311-321, 1992; Murray C J L. And Lopez A D. The globalburden of disease. Global burden of disease and injury series. Vol. 1.Cambridge, Mass.: Harvard University Press, 1996, p349-350; GlobalTuberculosis Programme: Anti-tuberculosis drug resistance, WHO Report1997: World Health Organization. 1997).

In 1950's, it was reported that non-tuberculosis mycobacteria (NTM) hasbeen able to cause diseases in human. After the report that M. aviumcomplex (MAC) would bring about systemic disease in the patients of AIDSin 1980, non-tuberculosis mycobacteria have taken an interest. Diseasescaused by non-tuberculosis mycobacteria are similar to tuberculosis inclinical condition and general pathological view. Non-tuberculosismycobacteria are distributed in a wide range of living environment, andit is difficult to judge whether they have pathogenicity or not inclinical test sample. Further, since they have resistance to a number ofdrugs for tuberculosis, the infection is hard to treat and therecurrence rate is high. The infection of non-tuberculosis mycobacteriashould be treated by other means than for tuberculosis, and therefore,accurate and fast method of detecting and identifying non-tuberculosismycobacteria is required. The accurate and fast method of detecting andidentifying both TB complex and NTM is also needed for effectivetreatment and management of tuberculosis.

Many a method has been developed to diagnose mycobacterial infection andto detect and identify mycobacteria strains. Among them, the followingmethods are used at present;

The first is a microbiological method, that is, smearing, staining andculturing test. However, this method is not suitable for mycobacteria,since they have long generation term and need long culturing time.Further, such pathogenic microorganism as mycobacteria is dangerous toinfect the personnel in culture room;

The second is a PCR (Polymerase Chain Reaction) method. It is highlysensitive and specific to the mycobacteria and very useful to detectmycobacteria which have a long culturing time. Especially, it does notrequire a culturing process but uses a small amount of DNA to beamplified, therefore, only a small amount of pathogens in test sample isenough to detect and identify mycobacteria. Many a PCR process has beenintroduced with different target DNAs each other, and IS6110 and 16SrRNA are often used as the target (Bauer J, Andersen A B, Kremer K, andMiorner H, Usefulness of spoligotyping to discriminate IS6110low-copy-number Mycobacterium tuberculosis complex strains cultured inDenmark, 1999, J. Clin. Microbiol. 37: 2602-2606; Troesch, A., H.Nguyen, C. G. Miyada, S. Desvarenne, T. R. Gingeras, P. M. Kaplan, P.Cros and C. Mabilat. 1999, Mycobacterium species identification andrifampin resistance testing with high-density DNA probe arrays, J. Clin.Microbiol. 37: 49-55);

The third is a physico-chemical process, in which lipid component inmycobacteria has been detected by HPLC, GC or mass spectrophotometer.This method is very specific but rquires expensive equipments;

The fourth is a method of detecting mycobacteria composition byserological method. This method uses a coagulation reaction of latexparticles or blood corpuscles adsorbed with antibody to mycobacterialantigen or enzyme-linked immunological method in which enzyme is linkedwith antibody. It is, however, very sensitive only to be proceededwithin a limited place. Further, it is difficult for this method todistinguish present infection from previous infection;

The next method to detect mycobacteria consists of infectingmycobacteria with mycobacteriophage L5 inserted with luciferase gene,and inspecting luminescence by luciferin in medium (W. R. Jacobs, R. G.Barletta, R. Udani, J. Chan, G. Kalkut, G. Sosne, T. Kieser, G. J.Sarkis, G. F. Hatful, and B. R. Bloom. 1993, Science 260: 819-822); and

The last is a method of detecting and identifying mycobacteria byhybridization of oligonucleotide (A. Troesch, H. Nguyen, C. G. Miyada,S. Desvarenne, T. R. Gingeras, P. M. Kaplan, P. Cros and C. Mabilat.1999. J. Clin. Microbiol 37: 49-55).

Besides Mycobacierium avium complex (MAC) described above, M. Fortuitum,M. chelonae complex, M. terrae and M. vaccae are also known asnon-tuberculosis mycobacteria. Among them, M. chelonae complex areclassified into M. chelonae and M. abscessus, and there is no means todistinguish one from the other at present.

SUMMARY OF THE INVENTION

To solve the problems in the prior method of detection andidentification of mycobacteria, it is an objective of the presentinvention to provide specific oligonucleotides as probes or primers forPCR which can be used to detect mycobacteria, to distinguish TB complexfrom NTM, and to identify species of mycobacteria with an accuracy andeffectiveness.

To accomplish the above objective, the present invention provides a DNAof ITS (Internal Transcribed Spacer region) of Mycobacterium fortuitum,Mycobacterium chelonae, Mycobacterium abscessus, Mycobacterium vaccae,Mycobacterium flavescens, Mycobacterium asiaticum, Mycobacteriumporcinum, Mycobacterium acapulcensis and Mycobacterium diernhoferi genesset forth in SEQ ID NOs: 1 to 9.

Further, the present invention provides, as a primer for PCR or a probefor hybridization, an oligonucleotide for detection of mycobacteria setin forth in one of SEQ ID NOs: 10 to 14;

an oligonucleotide for distinction of TB complex from NTB amongmycobacteria set in forth in one of SEQ ID NOs: 15 to 23;

an oligonucleotide for detection and identification of MAC(Mycobacterium avium and Mycobacterium intracellulare) set in forth inone of SEQ ID NOs: 24 to 27;

an oligonucleotide for detection and identification of Mycobacteriumfortuitum set in forth in one of SEQ ID NOs: 28 to 38;

an oligonucleotide for detection and identification of Mycobacteriumchelonae set in forth in one of SEQ ID NOs: 39 to 46;

an oligonucleotide for detection and identification of Mycobacteriumabscessus set in forth in one of SEQ ID NOs: 47 to 52;

an oligonucleotide for detection and identification of Mycobacteriumvaccae set in forth in one of SEQ ID NOs: 53 to 64;

an oligonucleotide for detection and identification of Mycobacteriumflavescens set in forth in one of SEQ ID NOs: 65 to 72;

an oligonucleotide for detection and identification of Mycobacteriumgordonae set in forth in one of SEQ ID NOs: 73 to 77;

an oligonucleotide for detection and identification of Mycobacteriumterrae set in forth in one of SEQ ID NOs: 78 to 100;

an oligonucleotide for detection and identification of Mycobacteriumscrofulaceum set in forth in one of SEQ ID NOs: 101 to 108;

an oligonucleotide for detection and identification of Mycobacteriumkansasii set in forth in one of SEQ ID NOs: 109 to 112;

an oligonucleotide for detection and identification of Mycobacteriumszulgai set in forth in one of SEQ ID NOs: 113 to 116;

an oligonucleotide for detection and identification of Mycobacteriummarinum and Mycobacterium ulcerans set in forth in one of SEQ ID NOs:117 to 119;

an oligonucleotide for detection and identification of Mycobacteriumgastri set in forth in one of SEQ ID NOs: 120 to 123;

an oligonucleotide for detection and identification of Mycobacteriumxenopi set in forth in one of SEQ ID NOs: 124 to 133;

an oligonucleotide for detection and identification of Mycobacteriumgenavense set in forth in one of SEQ ID NOs: 134 to 141;

an oligonucleotide for detection and identification of Mycobacteriummalmoense set in forth in one of SEQ ID NOs: 142 to 146;

an oligonucleotide for detection and identification of Mycobacteriumsimiae set in forth in one of SEQ ID NOs: 147 to 153;

an oligonucleotide for detection and identification of Mycobacteriumsmegmatis set in forth in one of SEQ ID NOs: 154 to 165;

an oligonucleotide for detection and identification of Mycobacteriumshimoidei set in forth in one of SEQ ID NOs: 166 to 172;

an oligonucleotide for detection and identification of Mycobacteriumhabana set in forth in one of SEQ ID NOs: 173 to 180;

an oligonucleotide for detection and identification of Mycobacteriumfarcinogen set in forth in one of SEQ ID NOs: 181 to 189;

an oligonucleotide for detection and identification of Mycobacteriumasiaticum set in forth in one of SEQ ID NOs: 190 to 193;

an oligonucleotide for detection and identification of Mycobacteriumporcinum set in forth in one of SEQ ID NOs: 194 to 205;

an oligonucleotide for detection and identification of Mycobacteriumacapulcensis set in forth in one of SEQ ID NOs: 206 to 215;

an oligonucleotide for detection and identification of Mycobacteriumdiernhoferi set in forth in one of SEQ ID NOs: 216 to 227;

an oligonucleotide for detection and identification of Mycobacteriumparatuberculosis set in forth in one of SEQ ID NOs: 228 to 240; and

an oligonucleotide for detection of Mycobacteria sp. set in forth in SEQID NO: 241.

In the prior method of detecting and identifying mycobacteria using PCR,only one or two strains can be detected. According to the presentinvention, however, almost all of mycobacteria strains can be detectedand identified, since primers and probes of the present invention havebeen designed form DNA sequences of ITS of mycobacteria. ITS has morepolymorphic region than 16S rRNA has and ITS also has conserved region,therefore, it is highly effective as a target DNA for distinction ofgenotype (Gurtler, V., and V. A. Stanisich, 1996, New approaches totyping and identification of bacteria using the 16S-23S rDNA spacerregion. Microbiol. 142: 3-16).

The inventors identified DNA sequences of ITS of non-tuberculosismycobacteria whose DNA had not yet been sequenced, such as Mycobacteriumfortuitum, Mycobacterium chelonae, Mycobacterium abscessus,Mycobacterium vaccae, Mycobacterium flavescens, Mycobacterium asiaticum,Mycobacterium porcinum, Mycobacterium acapulcensis and Mycobacteriumdiernhoferi. Using the DNA sequences, oligonucleotides for primers orprobes have been designed for detecting and identifying themycobacteria. Further, referring to the information on DNA sequence ofother mycobacteria disclosed in GenBank, and analyzing the informationwith multi-alignment and blast, distinctive regions of polymorphism wereselected to design oligonucleotides for primers or probes to detect andidentify mycobacteria. The oligonucleotides have been confirmed todetect and identify mycobacteria by specific hybridization andamplification with species-specific and genus-specific primers of PCR.

That is, the oligonucleotide probes of the present invention, attachedto solid substrate, are hybridized only with nucleotide sequence in ITSof specific mycobacteria, and therefore, they can detect and identifythe specific mycobacteria sensitively. Further, the oligonucleotideprimers of identical nucleotide sequence with the above probes can alsodetect and identify the specific mycobacteria by amplification in PCR.Using the oligonucleotide primers or probes made from ITS ofmycobacteria, it is possible to detect mycobacteria, distinguish TBcomplex from NTM, and identify mycobacteria species accurately andeffectively.

BRIEF DESCRIPTION OF THE DRAWINGS

The above and other objects, features, and advantages of the presentinvention will be apparent from the following detailed description ofthe preferred embodiments of the invention in conjunction with theaccompanying drawings, in which:

FIG. 1 is a schematic diagram showing ITS of mycobacteria and primersused to amplify the ITS by PCR;

FIG. 2 is a photograph showing the result of electrophoresis after PCRusing several mycobacteria strains and a pair of primers for ITSamplification including a part of 16S rRNA and 23S rRNA of mycobacteria;

FIG. 3 is a photograph showing the result of electrophoresis after PCRusing several mycobacteria strains and a pair of primers (ITSF and MYC2)for detecting mycobacteria;

FIG. 4 is a schematic diagram of multiplex PCR for detectingmycobacteria and simultaneously distinguishing TB complex from NTM;

FIG. 5 is a photograph showing the result of electrophoresis aftermultiplex PCR using a pair of primers (ITSF and MYC2) for detectingmycobacteria and a pair of primers (MTB2 and MYC2) for distinguishing TBcomplex from NTM;

FIG. 6 is a photograph showing the result of electrophoresis after PCRusing each mycobacteria and each pair of species-specific primersdesigned from nucleotide sequence of polymorphic region of each NTM; and

FIG. 7 is a photograph showing the result of electrophoresis after PCRusing several mycobacteria and each pair of species-specific primersdesigned from nucleotide sequence of polymorphic region of each NTM.

BEST MODE FOR CARRYING OUT THE INVENTION

Now, preferred embodiments of the present invention will be describedwith reference to the accompanying drawings.

EXAMPLE 1 Culture of Mycobacteria and Separation of Genome DNA

Standard strains of mycobacteria were obtained from KCTC (KoreanCollection for Type Culture) and ATCC (American Type CultureCollection), clinical strains were obtained from The Korean NationalTuberculosis Association, National Masan Tuberculosis Hospital and PusanNational University Hospital. The strains were stored and cultured underthe management of clinical microbiology specialist in Pusan NationalUniversity Hospital. The mycobacteria and pathogens used in the examplesare shown in Table 8.

Mycobacteria DNA was extracted by the following processes: Mycobacteriawere cultured on Ogawa medium. A loopful of cultured strain was put ineppendorf tube, mixed with 200 μl of InstaGene matrix (Bio-Rad Co.) andincubated at 56° C. for 30 minutes. After 10 minutes' vortex mixing, themixture was placed at 100° C. for 8 minutes. After another 10 minutes'vortex mixing, the mixture was centrifuged at 12,000 rpm for 3 minutes.The supematant was moved to another tube and stored at −20° C. 2 μl ofthe DNA solution of each strain was used in the succeeding PCR.

EXAMPLE 2 Manufacture of Primers for Amplifying ITS of Mycobacteria

The ITS of mycobacteria and primers used to amplify the ITS by PCR wereillustrated in FIG. 1. The primers for amplifying ITS of mycobacteriawere constructed from several section of conserved regions of 16S rRNAand 23S rRNA of mycobacteria. DNA sequences of 16S and 23S rRNA ofmycobacteria disclosed in GenBank were analyzed with multialignment andblast search. The primers were designed to amplify selectively about 500bp of ITS region with a part of 16S rRNA and 23S rRNA and have sequencesset in forth in SEQ ID NOs: 242 (ITSF) and 243 (ITSR). All the primersused in Examples were manufactured by the concentration of 50 nmol withPerkin-Elmer DNA Synthesizer by BioBasic (Canada).

EXAMPLE 3 PCR and Identification of Products

2 μl of the DNA solution obtained in Example 1 was used in PCR. Reactionsolution included: 500 mM KCl, 100 mM Tris HCl (pH 9.0), 1% TritonX-100, 0.2 mM dNTP (dATP, dGTP, dTTP and dCTP), 1.5 mM MgCl₂, 1 pmol ofprimer, 1 U of Taq DNA polymerase (Bio Basic Inc.). After denaturated at94° C. for 5 minutes, the solution was reacted 30 cycles of denaturationat 94° C. for 1 minute, annealing at 60° C. for 1 minute, and elongationat 72° C. for 1 minute. The samples were incubated further for 10minutes at 72° C. for complete elongation. After the reaction, the PCRproducts were identified by electrophoresis on 1.5% agarose gel. Asexpected from the information of GenBank, the ITS amplified using theconserved primers of 16S rRNA and 23S rRNA was about 500 bp.

FIG. 2 is a photograph showing the result of electrophoresis after PCRusing several mycobacteria strains and a pair of primers for ITSamplification including a part of 16S rRNA and 23S rRNA of mycobacteria.In this figure, M means molecular weight mark having intervals of 100bp, C is a control group, lanes 1 to 9 indicate, in turn, M. fortuitum,M. chelonae, M. intracellularae, M. avium, M. tuberculosis, M. agri, M.kansasii, M. gordonae, and M. tuberculosis H37Rv. It can be noted thatall mycobacteria of lanes 1 to 9 have amplification of ITS gene exceptcontrol group C.

EXAMPLE 4 Determination of DNA Sequence of Mycobacteria ITS

After the identification of PCR products, the reactants of M. fortuitum,M. chelonae, M. abscessus, M. vaccae, M. flavescens, M. asiaticum, M.porcinum, M. acapulcensis and M. diernhoferi, whose DNA sequences of ITShave not yet been determined, were amplified by PCR. The PCR productswere used directly in determining DNA sequence of ITS.

DNA was used by the concentration of 200 μmol. DNA sequence wasdetermined by dye terminator method using universal primer M13 with DNAauto sequencer (Perkin-Elmer, ABI prim 377 sequencer).

SEQ ID NOs 1 to 9 indicate DNA sequences of ITS of Mycobacteriumfortuitum, Mycobacterium chelonae, Mycobacierium abscessus,Mycobacterium vaccae, Mycobacterium flavescens, Mycobacterium asiaticum,Mycobacterium porcinum, Mycobacterium acapulcensis and Mycobacteriumdiernhoferi, in turn,

EXAMPLE 5 Design and Synthesis of Oligonucleotide Primer

Each primer of about 20 bp was designed from DNA sequences of ITS ofMycobacterium fortuitum, Mycobacterium cheloniae, Mycobacierizimabscessus, Mycobacterium vaccae, Mycobacterium flavescens, Mycobacteriumasiaticum, Mycobacterium porcinum, Mycobacterium acapulcensis andMycobacterium diernhoferi, obtained in Example 4, and compared with DNAsequences of ITS of other mycobacteria obtained from GenBank bymultialignment and blast.

Based on the result, conservative DNA sequences of mycobacteria ITS weredesigned as primers or probes for detection of mycobacteria, and DNAsequences of high polymorphism were designed as species-specific primersor probes. Tables 1 to 7 depict the designed primers or probes withtheir position and SEQ ID Nos.

TABLE 1 Target Strain Probe Name Position in ITS SEQ ID NO. MycobacteriaMYC1 Variabie to the 10 MYC2 species 11 MYC3 12 MYC4 13 MYC5 14 TBcomplex MTB1 166-185 15 MTB2 65-84 16 MTB3 20-39 17 MTB4 41-60 18 MTB560-79 19 MTB6  81-100 20 MTB7 125-144 21 MTB8 139-158 22 MTB9 203-222 23M. avium - M. MAC1 241-260 24 intracellularae MAC2 142-161 25 (MAC) MAC3 92-111 26 MAC4 117-136 27 M. fortuitum FOR1 40-59 28 FOR2 44-63 29 FOR364-83 30 FOR4 78-97 31 FOR5  89-108 32 FOR6 109-128 33 FOR7 114-133 34FOR8 134-153 35 FOR9 157-176 36 FOR10 246-265 37 FOR11 289-308 38 M.chelonae CHE1 11-30 39 CHE2 29-48 40 CHE3 58-77 41 CHE4 78-97 42 CHE5109-128 43 CHE6 132-151 44 CHE7 171-190 45 CHE8 246-265 46

TABLE 2 Target Strain Probe Name Position in ITS SEQ ID NO. M. abscessusABC1 37-56 47 ABC2 55-74 48 ABC3 247-266 49 ABC4 263-282 50 ABC5 270-28951 ABC6 261-280 52 M. vaccae VAC1 18-37 53 VAC2 38-57 54 VAC3 58-77 55VAC4 118-137 56 VAC5 138-157 57 VAC6 158-177 58 VAC7 178-197 59 VAC8199-218 60 VAC9 219-238 61 VAC10 265-284 62 VAC11 298-317 63 VAC12321-340 64 M. flavescens FLA1 12-31 65 FLA2 32-51 66 FLA3 52-71 67 FLA472-91 68 FLA5 105-124 69 FLA6 125-144 70 FLA7 173-192 71 FLA8 278-297 72M. gordonae GOR1  84-103 73 GOR2 249-268 74 GOR3 216-235 75 GOR4 201-22076 GOR5 223-242 77

TABLE 3 Target Strain Probe Name Position in ITS SEQ ID NO. M. terraeTER1 178-197 78 TER2 237-256 79 TER3 24-43 80 TER4 70-89 81 TER5  89-10882 TER6 102-121 83 TER7 122-141 84 TER8 142-161 85 TER9 162-181 86 TER10182-201 87 TER11 202-221 88 TER12 222-241 89 TER13 238-257 90 TER14307-326 91 TER15 322-341 92 TER16 342-361 93 TER17 362-381 94 TER18382-401 95 TER19 12-21 96 TER20 31-50 97 TER21 52-71 98 TER22 72-91 99TER23  82-101 100 M. scrofulaceum SCO1 118-137 101 SCO2 131-150 102 SCO3210-229 103 SCO4  84-103 104 SCO5 152-171 105 SCO6 200-219 106 SCO7221-240 107 SCO8 241-260 108 M. kansasii KAN1 35-54 109 KAN2 238-257 110KAN3  83-102 111 KAN4 214-233 112

TABLE 4 Target Strain Probe Name Position in ITS SEQ ID NO. M. szulgaiSZU1 124-143 113 SZU2 209-228 114 SZU3 227-246 115 SZU4 247-166 116 M.marinum and MAR-ULC1  85-104 117 M. ulcerans MAR-ULC2 128-147 118MAR-ULC3 224-243 119 M. gastri GAS1  85-104 120 GAS2 145-164 121 GAS3133-152 122 GAS4 239-258 123 M. xenopi XEN1 190-209 124 XEN2  1-20 125XEN3 21-40 126 XEN4 41-60 127 XEN5 61-80 128 XEN6  81-100 129 XEN7121-140 130 XEN8 141-160 131 XEN9 201-220 132 XEN10 221-240 133 M.genavense GEN1 190-209 134 GEN2  85-104 135 GEN3 131-150 136 GEN4147-166 137 GEN5 186-205 138 GEN6 206-225 139 GEN7 226-245 140 GEN8240-265 141 M. malmoense MAL1 203-222 142 MAL2 29-48 143 MAL3 136-155144 MAL4 222-241 145 MAL5 242-261 146 M. simiae SIM1  83-102 147 SIM2129-148 148 SIM3 208-227 149 SIM4 22-41 150 SIM5 80-99 151 SIM6 136-155152 SIM7 241-260 153

TABLE 5 Target Strain Probe Name Position in ITS SEQ ID NO. M. smegmatisSMEG1 17-36 154 SMEG2 37-56 155 SMEG3 57-76 156 SMEG4 77-96 157 SMEG5 97-116 158 SMEG6 117-136 159 SMEG7 137-156 160 SMEG8 157-176 161 SMEG9177-196 162 SMEG10 193-212 163 SMEG11 61-80 164 SMEG12 112-131 165 M.shimoidei SHI1  89-108 166 SHI2 20-39 167 SHI3 70-89 168 SH14  97-116169 SHI5 135-154 170 SHI6 224-243 171 SHI7 244-263 172 M. habana HAB1 86-105 173 HAB2 17-36 174 HAB3 51-70 175 HAB4  81-100 176 HAB5 134-153177 HAB6 175-194 178 HAB7 200-219 179 HAB8 242-261 180 M. farcinogenFAR1 122-141 181 FAR2 111-130 182 FAR3 22-41 183 FAR4 48-67 184 FAR576-95 185 FAR6 108-127 186 FAR7 114-133 187 FAR8 275-294 188 FAR9295-314 189

TABLE 6 Target Strain Probe Name Position in ITS SEQ ID NO. M. asiaticumASI1  82-101 190 ASI2 145-164 191 ASI3 189-208 192 ASI4 274-293 193 M.porcinum POR1 45-64 194 POR2 13-32 195 POR3 67-86 196 POR4  91-110 197POR5 115-134 198 POR6 137-156 199 POR7 164-183 200 POR8 194-213 201 POR9221-240 202 POR10 273-292 203 POR11 298-317 204 POR12 325-344 205 M.acapulcensis ACA1 66-85 206 ACA2 112-131 207 ACA3 132-151 208 ACA4178-197 209 ACA5 198-217 210 ACA6 219-238 211 ACA7 242-261 212 ACA8262-281 213 ACA9 318-337 214 ACA10 350-369 215 M. diernhoferi DIE1 16-35216 DIE2 36-55 217 DIE3 62-81 218 DIE4 103-122 219 DIE5 154-173 220 DIE6175-194 221 DIE7 195-214 222 DIE8 232-251 223 DIE9 261-280 224 DIE10282-301 225 DIE11 304-323 226 DIE12 344-363 227

TABLE 7 Target Strain Probe Name Position in ITS SEQ ID NO. M. para-PARA1  7-26 228 tuberculosis PARA2 30-49 229 PARA3 40-59 230 PARA4 50-69231 PARA5 71-90 232 PARA6  83-102 233 PARA7 103-122 234 PARA8 135-154235 PARA9 157-176 236 PARA10 178-197 237 PARA11 198-217 238 PARA12219-238 239 PARA13 241-260 240 M. sp SP1 225-244 241

EXAMPLE 6 Result of PCR Using Primers for Detecting Mycobacteria

Genus-specific primers, designed from conserved DNA sequence inmycobacteria were used for detecting mycobacteria. Among the primersmanufactured from the ITS sequence of 270-350 bp, a pair of primers,ITSF (SEQ ID NO. 242) and MYC2 (SEQ ID NO. 11) were used to proceed PCR.As a result, amplified nucleotides of about 350 bp were obtained inmycobacteria strains, while no amplification was occurred inStaphylococcus aureus, Enterococcus faecium and Serratia marcescens.Therefore, it is understood that the primers could be used for detectingmycobacteria.

FIG. 3 is a photograph showing the result of electrophoresis after PCRusing several mycobacteria strains and a pair of primers (ITSF and MYC2)for detecting mycobacteria. In this figure, M indicates a size marker of100 bp ladder; C indicates a negative control; lane 1 indicates M.abscessus ATCC 19977; lane 2, M. agri ATCC 27406; lane 3, M. asiaticumATCC 25276; lane 4, M. austroafricanum ATCC 33464; lane 5, M. avium ATCC25291; lane 6, M. bovis ATCC 19210; lane 7, M. chelonae ATCC 35752; lane8, M. flavescens ATCC 14474; lane 9, M. fortuitum ATCC 6841; lane 10, M.gordonae ATCC 14470; lane 11, M. intracellularae ATCC 13950; lane 12, M.kansasii ATCC 12478; lane 13, M. phlei ATCC 354; lane 14, M.scrofulaceutm ATCC 19981; lane 15, M. smegmatis ATCC 21701; lane 16, M.szulgai ATCC 35799; lane 17, M. terrae ATCC 15755; lane 18, M. trivialeATCC 23292; lane 19, M. tuberculosis H37Rv; and lane 20, M. vaccae ATCC15483. It can be seen that all mycobacteria of lanes 1 to 20 show ITSamplification except negative control C.

Table 8 shows the result of PCR using a pair of primers (ITSF and MYC2)for detecting mycobacteria. In this table, + indicates amplificationoccurred and − indicates no amplification. It can be seen that noamplification has occurred in other strains than mycobacteria.

TABLE 8 Name of strain result Name of strain result M. tuberculosisH37Rv + M. phlei ATCC 354 + M. bovis ATCC 19210 + Aeromonas hydrophila −M. avium ATCC 25291 + Burkholderia cepacia − M. intracellulare ATCC +Candida albicans − 13950 M. abscessus ATCC 19977 + Citrobacter freundii− M. chelonae ATCC 35752 + Enterobacter aerogenes − M. flavescens ATCC14474 + Enterobacter cloacae − M. fortuitum ATCC 6841 + Enterococcusfaecalis − M. gastri ATCC 15754 + Enterococcus faecium − M. genavenseATCC 51233 + Enterococcus raffinosis − M. gordonae ATCC 14470 +Escherichia coli − M. kansasii ATCC 12478 + Klebsiella pneumoniae − M.malmoense ATCC 29571 + Plesiomonas shigelloides − M. scrofulaceum ATCC +Proteus mirabilis − 19981 M. simiae ATCC 25275 + Proteus vulgaris − M.smegmatis ATCC 21701 + Providencia rettgeri − M. szulgai ATCC 35799 +Pseudomonas aeruginosa − M. terrae ATCC 15755 + Rahnella aquatilis − M.vaccae ATCC 15483 + Salmonella spp. − M. xenopi ATCC 19250 + Serratiamarcescens − M. marinum ATCC 927 + Shewanella putrefaciens − M.ulcerance ATCC 19423 + Shigella flexneri − M. porcinum ATCC 33776 +Shigella sonnei − M. asiaticum ATCC 25276 + Staphylococcus epidermidis −M. acapulcensis ATCC 14473 + Staphylococcus aureus − M. diernhoferi ATCC19340 + Streptococcus agalactiae − M. agri ATCC 27406 + Streptococcusintermidius − M. austroafricanum ATCC + Streptococcus pneumoniae − 33464M. triviale ATCC 23292 + Vibrio parahemolyticus −

EXAMPLE 7 Result of PCR Using Primers for TB Complex

Test for identifying each strain using the oligonucleotide primersmanufactured in the previous Examples was confirmed by amplification inPCR. For identifying TB complex, multiplex PCR was carried using a pairof primers (ITSF and MYC2) for detecting mycobacteria and a pair ofprimers (MTB2 and MYC2; SEQ ID Nos. 16 and 11) for distinguishing TBcomplex from NTM.

FIG. 4 is a schematic diagram of multiplex PCR for detectingmycobacteria and simultaneously distinguishing TB complex from NTM.

FIG. 5 is a photograph showing the result of electrophoresis aftermultiplex PCR using a pair of primers (ITSF and MYC2) for detectingmycobacteria and a pair of primers (MTB2 and MYC2) for distinguishing TBcomplex from NTM. In this figure, M indicates a size marker of 100 bpladder; C indicates a negative control; lanes 1 and 2 indicate TBcomplex, M. tuberculosis H37Rv and M. bovis, respectively; and lanes 3to 10 indicate M. avium, M. intracellularae, M. fortuitum, M. chelonae,M. gordonae, M. szulgai, M. terrae and M. scrofulaceum ATCC 19981, inturn. In lanes 1 and 2 of TB complex, two bands are formed by doubleamplification due to primers for detecting mycobacteria and primers forTB complex, while in the other lanes of NTM strains, only one band isformed, which confirms single amplification.

EXAMPLE 8 Result of PCR Using Primers for Identifying NTM Strains

PCR was carried out using species-specific primers manufactured from DNAsequence of polymorphic site of each strain for identifying NTM species.

Specifically, after selecting SEQ ID NOs. 16 and 21 for M. tuberculosisH37Rv and M. bovis, SEQ ID NOs. 24 and 27 for M. avium and M.intracellularae, SEQ ID NOs. 29 and 37 for M. fortuitum, SEQ ID NOs. 41and 44 for M. chelonae, SEQ ID NOs. 48 and 49 for M. abscessus, SEQ IDNOs. 55 and 63 for M. vaccae, SEQ ID NOs. 66 and 72 for M. flavescens,SEQ ID NOs. 73 and 75 for M. gordonae, SEQ ID NOs. 88 and 96 for M.terrae, SEQ ID NOs. 102 and 103 for M. scrofulaceum, SEQ ID NOs. 109 and110 for M. kansasii, SEQ ID NOs. 113 and 114 for M. szulgai, SEQ ID NOs.117 and 119 for M. marinum and M. ulcerans, SEQ ID NOs. 120 and 123 forM. gastri, SEQ ID NOs. 128 and 132 for M. xenopi, SEQ ID NOs. 135 and141 for M. genavense, SEQ ID NOs. 143 and 145 for M. malmoense, SEQ IDNOs. 147 and 149 for M. simiae, and SEQ ID NOs. 154 and 159 for M.smegmalis, each mycobacteria was carried out PCR using each pair ofprimers of which the first has been sense strand and the second has beenantisense strand. After the reaction, each resultant was treated byelectrophoresis.

FIG. 6 is a photograph showing the result of electrophoresis after PCRusing each mycobacteria and each pair of species-specific primersdesigned from nucleotide sequence of polymorphic region of each NTM. Inthis figure, M indicates a size marker of 100 bp ladder; C indicates anegative control; lanes 1 and 2 indicate TB complex, M. tuberculosisH37Rv and M. bovis ATCC 19210, respectively; lanes 3 and 4 indicate M.avium ATCC 25291 and M. intracellularae ATCC 13950, respectively; lane 5indicates M. fortuitum ATCC 6841; lane 6, M. chelonae ATCC 35752; lane7, M. abscessus ATCC 19977; lane 8, M. vaccae ATCC 15483; lane 9, M.flavescens ATCC 14474; lane 10, M. gordonae ATCC 14470; lane 11, M.terrae ATCC 15755; lane 12, M. scrofulaceum ATCC 19981; lane 13, M.kansasii ATCC 12478; lane 14, M. szulgai ATCC 35799; lane 15, M. marinumATCC 927; lane 16, M. ulcerans ATCC 19423; lane 17, M. gastri ATCC15754; lane 18, M. xenopi ATCC 19250; lane 19, M. genavense ATCC 1233;lane 20, M. malmoense ATCC 29571; lane 21, M. simiae ATCC 25275; andlane 22 indicates M. smegmatis ATCC 21701. Species-specificamplifications can be seen in lanes 1 to 22.

FIG. 7 is a photograph showing the result of electrophoresis after PCRusing several mycobacteria and each pair of species-specific primersdesigned from nucleotide sequence of polymorphic region of each NTM. Inthis figure, the first photograph using the pair of primers (ITSF andMYC2) specific for TB complex shows amplifications in lanes 1 and 2 ofM. tuberculosis H37Rv and M. bovis; the second using the pair of primers(MAC1 and MAC4) specific for M. avium and M. intracellularae showsamplifications in lanes 3 and 4 of M. avium and M. intracellularae; thethird using the pair of primers (FOR2 and FOR10) specific for M.fortuitum shows amplification in lane 5 of M. fortuitum; the fourthusing the pair of primers (CHE3 and CHE6) specific for M. chelotiaeshows amplification in lane 6 of M. chelonae; the fifth using the pairof primers (GOR1 and GOR2) specific for M. gordonae shows amplificationin lane 7 of M. gordonae; the sixth using the pair of primers (SZU1 andSZU2) specific for M. szulgai shows amplification in lane 8 of M.szulgai; and the seventh using the pair of primers (SCO1 and SCO2)specific for M. scrofulaceum shows amplification in lane 10 of M.scrofulaceum. Lane 9 indicates M. terrae, which shows no amplificationwith the above species—specific primer pairs.

Therefore, PCR using each pair of species-specific primers can detectand identify specifically each species of mycobacteria.

INDUSTRIAL APPLICABILITY

As described above, identifying DNA sequences of ITS (InternalTranscribed Spacer region) of non-tuberculosis mycobacteria,Mycobacterium fortuitum, Mycobacterium chelonae, Mycobacteriumabscessus, Mycobacterium vaccae, Mycobacterium flavescens, Mycobacteriumasiaticum, Mycobacterium porcinur, Mycobacterium acapulcensis andMycobacterium diernhoferi, and using the DNA sequences, oligonucleotideprimers or probes can been designed for detecting and identifyingmycobacteria. Using the primers for PCR or probes for hybridization, itis possible to detect mycobacteria, distinguish TB complex from NTM. andidentify mycobacteria species with rapidity and effectiveness. Suchdetection and identification method makes the diagnosis of complexinfection effective, and therefore, it is possible to treat accuratelymycobacterial infection including tuberculosis.

While this invention has been described in connection with what ispresently considered to be the most practical and preferred embodiments,it is to be understood that the invention is not limited to thedisclosed embodiment, but, on the contrary, it is intended to covervarious modifications and equivalent arrangements included within thespirit and scope of the appended claims.

SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 243 <210> SEQ ID NO 1 <211>LENGTH: 386 <212> TYPE: DNA <213> ORGANISM: Mycobacterium fortuitum<400> SEQUENCE: 1 aggagcacca cgaaaagggt tgagacactg ggtcttaccc gagccgtgaggaaccggttg 60 cctgtagtgg gcacggtttg gtgcacaaca aacttttttg actgccagacacactattgg 120 gctttgagac aacaggcccg tgcccctttt ggggggtggc atccggttgcgggtgtcggc 180 gtgttgttgc ctcactttgg tggtggggtg tggtgtttga tttgtggatagtggttgcga 240 gcatctagca cgcagaatcg tgtggtctca ctccttgtgg gtggggctggttttgtgtgt 300 tgatgtgcaa tttcttttga aactcatttt tggtttttgt gttgtaagtgtttaagggcg 360 catggtggat gccttggcag gatcca 386 <210> SEQ ID NO 2 <211>LENGTH: 338 <212> TYPE: DNA <213> ORGANISM: Mycobacterium chelonae <400>SEQUENCE: 2 aggagcacca tttcccagcc gaatgagctt gggaacataa agcgagtttctgtagtggtt 60 actcgcttgg tgaatatgtt ttataaatcc tgtccacccc gtggataggtagtcggcaaa 120 acgtcggact gtcaatagaa ttgaaacgct ggcacactgt tgggtcctgaggcaacacat 180 tgtgttgtca ccctgcttgg tggtggggtg tggtctttga cttatggatagtggttgcga 240 gcatctaaca aacctcgctc gtttacgagt gaggttagtt tttgcaatttattagctaag 300 ttcttaaggg cacatggtgg atgccttggc aggatcca 338 <210> SEQID NO 3 <211> LENGTH: 317 <212> TYPE: DNA <213> ORGANISM: Mycobacteriumabscessus <400> SEQUENCE: 3 aaggagcacc atttcccagt cgaatgaact agggaacataaagtaggcat ctgtagtgga 60 tatctacttg gtgaatatgt tttgtaaatc ctgtccaccccgtggatggg tagtcggcaa 120 aacgtcggac tgtcataaga attgaaacgc tggcacactgttgggtcccg aggcaacacg 180 ttgtgttgtc accctgcttg gtggtggggt gtggactttgacttctgaat agtggttgcg 240 agcatctaaa catagcctcg ctcgttttcg agtggggctggtttttgcaa ttttattagc 300 taagttctta agggcgc 317 <210> SEQ ID NO 4 <211>LENGTH: 429 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 4 aaggagcacc acgagaatca ggcccgccca catcgtgtgg gggttcggtgatctgatcga 60 ttcgttggat ggcctttcat ctgtagtgga tgggggtctg gtgcacatgacaaacttggc 120 cgagccggta gggaatgccg gcgagggaaa tcatcagaca cactattgggctttgagaca 180 acaagcccgt gccccttttt ttgggggtgg ctctgcgttg gcagggtcggcgtgttgttg 240 ccccgctttg gtggtggggt gtggtgtttg attcgtggat agtggttgcgagcatctgaa 300 tgcacagcgc ttgtggtgtt gtgtgttcgg tgtaatgcaa atttttctgatactcgcatg 360 cagtcccttt ttgggggtgt gtgtgggtga ctcatttttt ggttttgtgttgtaagtgtt 420 taagggcgc 429 <210> SEQ ID NO 5 <211> LENGTH: 379 <212>TYPE: DNA <213> ORGANISM: Mycobacterium flavescens <400> SEQUENCE: 5aaggagcacc atttattgtt cccccgtccc cacgtgtgtg ggatcagtgc ggttgggaga 60tcagtgccgg gcctgtagtg ggtttccggt gggtgcacaa caaacgtgag aagtggtgtg 120ggaacactgc tttgaggaat catcagacac actattgggc tttgaggcaa caggcccgt 180gtttccctgg ccactgtgtg tggtgggggg tctggtgtcg ccctgtcttt ggtggtgggg 240tgtggtgttt gattcgtgga tagtggttgc gagcatctga acaggtggct cccttttggg 300ggttgcttgt tttgataatg caatttttta ttcttccgag aatatttttc tgttttgtgt 360tgtaagtgtt taagggcgc 379 <210> SEQ ID NO 6 <211> LENGTH: 364 <212> TYPE:DNA <213> ORGANISM: Mycobacterium asiaticum <400> SEQUENCE: 6 tggatccgacgaagtcgtaa caaggtagcc gtaccggaag gtgcggctgg atcacctcct 60 ttctaaggagcaccacgaaa gcatcccaat tggtgggatg caggccgtgt ggagttctcg 120 tctgtagtggacgagggctg ggtgcacaac aacaagcaag ccagacacac tattgggtcc 180 tgagacaacactcgggcgct agcacgaagt gttgtccctc catcttggtg gtggggtgtg 240 gtgtttgagaactggatagt ggttgcgagc atcaactgat cgcgtcgccg ttcgcggtgg 300 cgtgttcttttgtgcaattt taaattcttt ggtttttgta gtgtttgtaa gtgtctaagg 360 gcgc 364<210> SEQ ID NO 7 <211> LENGTH: 402 <212> TYPE: DNA <213> ORGANISM:Mycobacterium porcinum <400> SEQUENCE: 7 aaggagcacc gattcgattcccccgccgtc cgcgtagtcg tgggcagtag tgcggttggg 60 atatttcagc caggacctgtagtgggtgtc tggtgggtgc aaatgacaaa cgttgagccg 120 gcgcgggaaa gcgttggtgatggaactgct gaacacacta ttgggctttg agacaacagg 180 cccgtgcccc tttcggggggtggcattccg ttgggagtgt cggcgtgttg ttgctccgct 240 ttggtggtgg ggtgtggtgtttgatttgtg gatagtggtt gcgagcatct agcacgcagt 300 gtggctgggg gccttcgggtttccggtctt gttgtgtgtt gatgtgcaat ttcttttgaa 360 actcattttt ggtttttgtgttgtaagtgt ttaagggcgc at 402 <210> SEQ ID NO 8 <211> LENGTH: 459 <212>TYPE: DNA <213> ORGANISM: Mycobacterium acapulcensis <400> SEQUENCE: 8aaggagcacc agttattttc ccccgccccc acgcctgtgg gatcagttgc ggttgggata 60tttagtgcca gcacctgtag tgggtgtttc ggccggtgca cagcaaacgt tgatcgtctg 120gtggggaaag ccgggcgtga aagattgcca gacacactat tgggctttga gacaacaagc 180ccgtcgcctc tttgtcccga gtgtgggata tcggagaagg agcaccacga gacctgttgc 240ccgcccacat cgtgtgggag ttcggtgact caggcgattc gggggttggt tgtggttgtc 300gccctgcttt ggtggtgggg tgtggtgttt gatttgtgga tagtggttgc gagcatctga 360acgcagagac ctgtgtgggt ttttgtgttc tgataatgca atttttattc ttccgagaat 420attttttgat ctgttttgtg tgtaagtgtt taagggcgc 459 <210> SEQ ID NO 9 <211>LENGTH: 418 <212> TYPE: DNA <213> ORGANISM: Mycobacterium diernhoferi<400> SEQUENCE: 9 aaggagcacc acgagacctg ttgcccgccc acatcgtgtg ggagttcggtgactcaggcg 60 attcgttgga tggcctcaca cctgtagtgg gtgggggtct ggtgcacaacaaactttgag 120 aaactgccag acacactatt gggctttgag acaacaggcc ctgcggtgccggactcgttg 180 gagtcctggt tgccggccgc gagtcccgga agcgattctg gttcggacggtgtctgttgt 240 tgctccatct ttggtggtgg ggtgtggtgt ttgatttgtg gatagtggttgcgagcatct 300 agcacgcaag aggagtctgg gtttccttcg ggagcccggg tttttgttgtgtgtgtttga 360 tgtgcaattt ttttcttcta tttggtttta tctgtgttgt aagtgtttaagggcgcat 418 <210> SEQ ID NO 10 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacteria <400> SEQUENCE:10 tggtggggtg tggtgtttga 20 <210> SEQ ID NO 11 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteria<400> SEQUENCE: 11 tggatagtgg ttgcgagcat 20 <210> SEQ ID NO 12 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacteria <400> SEQUENCE: 12 ccatcttggt ggtggggtgt 20 <210>SEQ ID NO 13 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacteria <400> SEQUENCE: 13 cacactattgggccctgagg 20 <210> SEQ ID NO 14 <211> LENGTH: 10 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacteria <400> SEQUENCE:14 aaggagcacc 10 <210> SEQ ID NO 15 <211> LENGTH: 20 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting TB complex <400>SEQUENCE: 15 cactcggact tgttccaggt 20 <210> SEQ ID NO 16 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detecting TBcomplex <400> SEQUENCE: 16 tggtggggcg taggccgtga 20 <210> SEQ ID NO 17<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting TB complex <400> SEQUENCE: 17 ccccaactgg tggggcgtag 20 <210>SEQ ID NO 18 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting TB complex <400> SEQUENCE: 18 ccgtgagggg ttcttgtctg20 <210> SEQ ID NO 19 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting TB complex <400> SEQUENCE: 19 gtagtgggcgagagccgggt 20 <210> SEQ ID NO 20 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting TB complex <400> SEQUENCE: 20catgacaaca aagttggcca 20 <210> SEQ ID NO 21 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting TB complex <400>SEQUENCE: 21 caacactcgg acttgttcca 20 <210> SEQ ID NO 22 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detecting TBcomplex <400> SEQUENCE: 22 gttccaggtg ttgtcccacc 20 <210> SEQ ID NO 23<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting TB complex <400> SEQUENCE: 23 cgagcatcaa tggatacgct 20 <210>SEQ ID NO 24 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium avium complex(MAC) <400> SEQUENCE: 24gttcatcgaa atgtgtaatt 20 <210> SEQ ID NO 25 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumavium complex(MAC) <400> SEQUENCE: 25 gtgtggagtc cctccatctt 20 <210> SEQID NO 26 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium avium complex(MAC) <400> SEQUENCE: 26aaatgattgc cagacacact 20 <210> SEQ ID NO 27 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumavium complex(MAC) <400> SEQUENCE: 27 ccctgagaca acactcggtc 20 <210> SEQID NO 28 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium fortuitum <400> SEQUENCE: 28cgagccgtga ggaaccggtt 20 <210> SEQ ID NO 29 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumfortuitum <400> SEQUENCE: 29 ccgtgaggaa ccggttgcct 20 <210> SEQ ID NO 30<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium fortuitum <400> SEQUENCE: 30 gtagtgggcacggtttggtg 20 <210> SEQ ID NO 31 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium fortuitum <400>SEQUENCE: 31 ttggtgcaca acaaactttt 20 <210> SEQ ID NO 32 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium fortuitum <400> SEQUENCE: 32 caaacttttt tgactgccag 20<210> SEQ ID NO 33 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium fortuitum <400> SEQUENCE: 33acacactatt gggctttgag 20 <210> SEQ ID NO 34 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumfortuitum <400> SEQUENCE: 34 ctattgggct ttgagacaac 20 <210> SEQ ID NO 35<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium fortuitum <400> SEQUENCE: 35 aggcccgtgccccttttggg 20 <210> SEQ ID NO 36 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium fortuitum <400>SEQUENCE: 36 tggcatccgg ttgcgggtgt 20 <210> SEQ ID NO 37 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium fortuitum <400> SEQUENCE: 37 tagcacgcag aatcgtgtgg 20<210> SEQ ID NO 38 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium fortuitum <400> SEQUENCE: 38ggttttgtgt gttgatgtgc 20 <210> SEQ ID NO 39 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumchelonae <400> SEQUENCE: 39 tttcccagcc gaatgagctt 20 <210> SEQ ID NO 40<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium chelonae <400> SEQUENCE: 40 ttgggaacataaagcgagtt 20 <210> SEQ ID NO 41 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium chelonae <400>SEQUENCE: 41 gttactcgct tggtgaatat 20 <210> SEQ ID NO 42 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium chelonae <400> SEQUENCE: 42 gttttataaa tcctgtccac 20 <210>SEQ ID NO 43 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium chelonae <400> SEQUENCE: 43gtagtcggca aaacgtcgga 20 <210> SEQ ID NO 44 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumchelonae <400> SEQUENCE: 44 tcaatagaat tgaaacgctg 20 <210> SEQ ID NO 45<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium chelonae <400> SEQUENCE: 45 ggcaacacattgtgttgtca 20 <210> SEQ ID NO 46 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium chelonae <400>SEQUENCE: 46 taacaaacct cgctcgttta 20 <210> SEQ ID NO 47 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium abscessus <400> SEQUENCE: 47 cataaagtag gcatctgtag 20<210> SEQ ID NO 48 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium abscessus <400> SEQUENCE: 48agtggatatc tacttggtga 20 <210> SEQ ID NO 49 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumabscessus <400> SEQUENCE: 49 taaacatagc ctcgctcgtt 20 <210> SEQ ID NO 50<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium abscessus <400> SEQUENCE: 50 cgttttcgagtggggctggt 20 <210> SEQ ID NO 51 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium abscessus <400>SEQUENCE: 51 gagtggggct ggtttttgca 20 <210> SEQ ID NO 52 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium abscessus <400> SEQUENCE: 52 ctcgttttcg agtggggctg 20<210> SEQ ID NO 53 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium vaccae <400> SEQUENCE: 53tcaggcccgc ccacatcgtg 20 <210> SEQ ID NO 54 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumvaccae <400> SEQUENCE: 54 tgggggttcg gtgatctgat 20 <210> SEQ ID NO 55<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium vaccae <400> SEQUENCE: 55 cgattcgttg gatggccttt20 <210> SEQ ID NO 56 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium vaccae <400> SEQUENCE: 56ggccgagccg gtagggaatg 20 <210> SEQ ID NO 57 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumvaccae <400> SEQUENCE: 57 ccggcgaggg aaatcatcag 20 <210> SEQ ID NO 58<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium vaccae <400> SEQUENCE: 58 acacactatt gggctttgag20 <210> SEQ ID NO 59 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium vaccae <400> SEQUENCE: 59acaacaagcc cgtgcccctt 20 <210> SEQ ID NO 60 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumvaccae <400> SEQUENCE: 60 ttttgggggt ggctctgcgt 20 <210> SEQ ID NO 61<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium vaccae <400> SEQUENCE: 61 tggcagggtc ggcgtgttgt20 <210> SEQ ID NO 62 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium vaccae <400> SEQUENCE: 62tgtttgattc gtggatagtg 20 <210> SEQ ID NO 63 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumvaccae <400> SEQUENCE: 63 gaatgcacag cgcttgtggt 20 <210> SEQ ID NO 64<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium vaccae <400> SEQUENCE: 64 gtgtgttcgg tgtaatgcaa20 <210> SEQ ID NO 65 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium flavescens <400> SEQUENCE:65 tttattgttc ccccgtcccc 20 <210> SEQ ID NO 66 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumflavescens <400> SEQUENCE: 66 acgtgtgtgg gatcagtgcg 20 <210> SEQ ID NO67 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium flavescens <400> SEQUENCE: 67 gttgggagatcagtgccggg 20 <210> SEQ ID NO 68 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium flavescens <400>SEQUENCE: 68 cctgtagtgg gtttccggtg 20 <210> SEQ ID NO 69 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium flavescens <400> SEQUENCE: 69 cgtgagaagt ggtgtgggaa 20<210> SEQ ID NO 70 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium flavescens <400> SEQUENCE:70 cactgctttg aggaatcatc 20 <210> SEQ ID NO 71 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumflavescens <400> SEQUENCE: 71 ggcccgttgt ttccctggcc 20 <210> SEQ ID NO72 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium flavescens <400> SEQUENCE: 72 tgaacaggtggctccctttt 20 <210> SEQ ID NO 73 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium gordonae <400>SEQUENCE: 73 cgacaacaag ctaagccaga 20 <210> SEQ ID NO 74 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium gordonae <400> SEQUENCE: 74 tgttcttttt gtgcaatttt 20 <210>SEQ ID NO 75 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium gordonae <400> SEQUENCE: 75gcatcaaaat gtatgcgttg 20 <210> SEQ ID NO 76 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumgordonae <400> SEQUENCE: 76 aaaatgtatg cgttgtcgtt 20 <210> SEQ ID NO 77<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium gordonae <400> SEQUENCE: 77 cggcaacgtgttctttttgt 20 <210> SEQ ID NO 78 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 78 ttgagttgtg gatagtggtt 20 <210> SEQ ID NO 79 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 79 gtcgagtgtt tagagagtaa 20 <210>SEQ ID NO 80 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium terrae <400> SEQUENCE: 80 cgcactgggcgcattccgag 20 <210> SEQ ID NO 81 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 81 gtgggggctg ggtgcacaac 20 <210> SEQ ID NO 82 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 82 caacgttgaa aacaagatcg 20 <210>SEQ ID NO 83 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium terrae <400> SEQUENCE: 83 aattacgaacaacaacaagc 20 <210> SEQ ID NO 84 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 84 ttgcgagatc atcaactgcc 20 <210> SEQ ID NO 85 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 85 aggagtcctt gggggtttct 20 <210>SEQ ID NO 86 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium terrae <400> SEQUENCE: 86 ggtggccggcttttgtgctg 20 <210> SEQ ID NO 87 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 87 ggcacactgt tgggtcctga 20 <210> SEQ ID NO 88 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 88 ggcaacaggc ccgtttgtgc 20 <210>SEQ ID NO 89 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium terrae <400> SEQUENCE: 89 ccccgggtgggggtgggtgt 20 <210> SEQ ID NO 90 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 90 gtgttgttgt cgcctcacac 20 <210> SEQ ID NO 91 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 91 taacaagcag atttttggtc 20 <210>SEQ ID NO 92 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium terrae <400> SEQUENCE: 92 tggtctgtttgttttgcaat 20 <210> SEQ ID NO 93 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 93 ttttgtttct tggtttttgt 20 <210> SEQ ID NO 94 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 94 gtttgtaagt gtttaagggc 20 <210>SEQ ID NO 95 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium terrae <400> SEQUENCE: 95 gcatggtggatgccttggca 20 <210> SEQ ID NO 96 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 96 ctttttcccc cgtgcctcac 20 <210> SEQ ID NO 97 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 97 atgggtgagg gtttttgcgg 20 <210>SEQ ID NO 98 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium terrae <400> SEQUENCE: 98 ttgggacagtgtttgccggt 20 <210> SEQ ID NO 99 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium terrae <400>SEQUENCE: 99 gcctgtagtg ggtggccggt 20 <210> SEQ ID NO 100 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium terrae <400> SEQUENCE: 100 ggtggccggt ggtgcagagg 20 <210>SEQ ID NO 101 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium scrofluceum <400> SEQUENCE:101 cctgaggcaa cactcggctc 20 <210> SEQ ID NO 102 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumscrofluceum <400> SEQUENCE: 102 tcggctcgtt ctgagtggtg 20 <210> SEQ ID NO103 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium scrofluceum <400> SEQUENCE: 103 taaacggatgcgtggccgaa 20 <210> SEQ ID NO 104 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium scrofluceum<400> SEQUENCE: 104 caacagcaaa tgattgccag 20 <210> SEQ ID NO 105 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium scrofluceum <400> SEQUENCE: 105 ccctccatcttggtggtggg 20 <210> SEQ ID NO 106 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium scrofluceum<400> SEQUENCE: 106 tgcgagcatc taaacggatg 20 <210> SEQ ID NO 107 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium scrofluceum <400> SEQUENCE: 107 gtggccgaacggtggcgtgt 20 <210> SEQ ID NO 108 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium scrofluceum<400> SEQUENCE: 108 tcgttgaaat gtgtaatttc 20 <210> SEQ ID NO 109 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium ka <400> SEQUENCE: 109 gtggggtgca agccgtgagg 20<210> SEQ ID NO 110 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium kansasii <400> SEQUENCE: 110gcgtgttctt ttgtgcaatt 20 <210> SEQ ID NO 111 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumkansasii <400> SEQUENCE: 111 gcaactgtaa atgaatcacc 20 <210> SEQ ID NO112 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium kansasii <400> SEQUENCE: 112 ctggatgcgctgccgttcgt 20 <210> SEQ ID NO 113 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium szulgai <400>SEQUENCE: 113 aacactcagg cttggccaga 20 <210> SEQ ID NO 114 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium szulgai <400> SEQUENCE: 114 caattggatg cgctgccctc 20 <210>SEQ ID NO 115 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium szulgai <400> SEQUENCE: 115tcgtggtggc gtgttctttt 20 <210> SEQ ID NO 116 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumszulgai <400> SEQUENCE: 116 gtgcaatttt aattctttgg 20 <210> SEQ ID NO 117<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium marinum and Mycobacterium ulcerans <400>SEQUENCE: 117 aacaacaagc aagccagaca 20 <210> SEQ ID NO 118 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium marinum and Mycobacterium ulcerans <400> SEQUENCE: 118atctctgttg gtttcgggat 20 <210> SEQ ID NO 119 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriummarinum and Mycobacterium ulcerans <400> SEQUENCE: 119 ccttttggtggcgtgttctg 20 <210> SEQ ID NO 120 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium gastri <400>SEQUENCE: 120 aacagcaagc aagccagaca 20 <210> SEQ ID NO 121 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium gastri <400> SEQUENCE: 121 ctcgtccaag agtgttgtcc 20 <210>SEQ ID NO 122 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium gastri <400> SEQUENCE: 122gcttgtcttg gactcgtcca 20 <210> SEQ ID NO 123 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumgastri <400> SEQUENCE: 123 cagggtagcg tgttcttttg 20 <210> SEQ ID NO 124<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium xenopi <400> SEQUENCE: 124 catctggcaa agactgtggt20 <210> SEQ ID NO 125 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium xenopi <400> SEQUENCE: 125agggagcacc gtaaacgcat 20 <210> SEQ ID NO 126 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumxenopi <400> SEQUENCE: 126 cccgcgtggg gtggggtgtg 20 <210> SEQ ID NO 127<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium xenopi <400> SEQUENCE: 127 ggttcggcgt gttgtggcgt20 <210> SEQ ID NO 128 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium xenopi <400> SEQUENCE: 128cgggccgagg tgttgggcag 20 <210> SEQ ID NO 129 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumxenopi <400> SEQUENCE: 129 caggcagtaa ccgccggcac 20 <210> SEQ ID NO 130<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium xenopi <400> SEQUENCE: 130 caggcagtaa ccgccggcac20 <210> SEQ ID NO 131 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium xenopi <400> SEQUENCE: 131tccgcgtggt ggcggggtgt 20 <210> SEQ ID NO 132 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumxenopi <400> SEQUENCE: 132 agactgtggt aagcggtttt 20 <210> SEQ ID NO 133<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium xenopi <400> SEQUENCE: 133 tgttgagtgt tttctggtgt20 <210> SEQ ID NO 134 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium genavense <400> SEQUENCE:134 cattgaatag tggttgcgag 20 <210> SEQ ID NO 135 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumgenavense <400> SEQUENCE: 135 aacaacaggc aatcgccgga 20 <210> SEQ ID NO136 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium genavense <400> SEQUENCE: 136 tcggccgactgaggtcgacg 20 <210> SEQ ID NO 137 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium genavense <400>SEQUENCE: 137 gacgtggtgt ccctccatct 20 <210> SEQ ID NO 138 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium genavense <400> SEQUENCE: 138 ttgagcattg aatagtggtt 20<210> SEQ ID NO 139 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium genavense <400> SEQUENCE:139 gcgagcatct agacggatgc 20 <210> SEQ ID NO 140 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumgenavense <400> SEQUENCE: 140 gttccccagt ggtgcgcgtt 20 <210> SEQ ID NO141 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium genavense <400> SEQUENCE: 141 cgtcaaaaatgtgtaatttt 20 <210> SEQ ID NO 142 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium malmoense <400>SEQUENCE: 142 agcatctaaa cggatgcgct 20 <210> SEQ ID NO 143 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium malmoense <400> SEQUENCE: 143 tggtggggtg caagccgtga 20<210> SEQ ID NO 144 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium malmoense <400> SEQUENCE:144 ccagtccgcg tggtgtcccc 20 <210> SEQ ID NO 145 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriummalmoense <400> SEQUENCE: 145 tgcccgtaga cgcgtattcg 20 <210> SEQ ID NO146 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium malmoense <400> SEQUENCE: 146 tttgtgtaatttcttctttg 20 <210> SEQ ID NO 147 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium simiae <400>SEQUENCE: 147 acaacaacag gcaatcgcca 20 <210> SEQ ID NO 148 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium simiae <400> SEQUENCE: 148 actcggccga cttcggttga 20 <210>SEQ ID NO 149 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium simiae <400> SEQUENCE: 149gagcatctaa atgaacgcgt 20 <210> SEQ ID NO 150 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumsimiae <400> SEQUENCE: 150 tccaattggt ggggtgtgag 20 <210> SEQ ID NO 151<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium simiae <400> SEQUENCE: 151 tgcacaacaa caggcaatcg20 <210> SEQ ID NO 152 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium simiae <400> SEQUENCE: 152cgacttcggt tgaagtggtg 20 <210> SEQ ID NO 153 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumsimiae <400> SEQUENCE: 153 tacgtgttcg ttttgtgtaa 20 <210> SEQ ID NO 154<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium smegmatis <400> SEQUENCE: 154 gacactctccgttggggagg 20 <210> SEQ ID NO 155 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium smegmatis <400>SEQUENCE: 155 gtgtgagccg tgaggagctg 20 <210> SEQ ID NO 156 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium smegmatis <400> SEQUENCE: 156 gagcgctgta gtggcgccgg 20<210> SEQ ID NO 157 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium smegmatis <400> SEQUENCE:157 cttggtgcac agcaaacgtt 20 <210> SEQ ID NO 158 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumsmegmatis <400> SEQUENCE: 158 gagatgcggt gtgggaaacg 20 <210> SEQ ID NO159 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium smegmatis <400> SEQUENCE: 159 ctgtttcgatggactgccag 20 <210> SEQ ID NO 160 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium smegmatis <400>SEQUENCE: 160 acacactatt gggccctgag 20 <210> SEQ ID NO 161 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium smegmatis <400> SEQUENCE: 161 acaacaggcc cgcgttcccg 20<210> SEQ ID NO 162 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium smegmatis <400> SEQUENCE:162 tcccgttggg ggcggggggt 20 <210> SEQ ID NO 163 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumsmegmatis <400> SEQUENCE: 163 gggtgtgttg ttgccctgct 20 <210> SEQ ID NO164 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium smegmatis <400> SEQUENCE: 164 gctgtagtggcgccggcttg 20 <210> SEQ ID NO 165 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium smegmatis <400>SEQUENCE: 165 aaacgctgtt tcgatggact 20 <210> SEQ ID NO 166 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium shimoidei <400> SEQUENCE: 166 aacaacaagc gagaagccga 20<210> SEQ ID NO 167 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium shimoidei <400> SEQUENCE:167 ggcttccgca gtgggcggaa 20 <210> SEQ ID NO 168 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumshimoidei <400> SEQUENCE: 168 gggcgcgggc tgggtgcaca 20 <210> SEQ ID NO169 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium shimoidei <400> SEQUENCE: 169 gcgagaagccgagcacactg 20 <210> SEQ ID NO 170 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium shimoidei <400>SEQUENCE: 170 cccgggccct ttggggttgg 20 <210> SEQ ID NO 171 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium shimoidei <400> SEQUENCE: 171 aagcaaaact tggttgtttt 20<210> SEQ ID NO 172 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium shimoidei <400> SEQUENCE:172 gtttgtcgag ttgttttctt 20 <210> SEQ ID NO 173 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumhabana <400> SEQUENCE: 173 aacaacaggc aatcgccaga 20 <210> SEQ ID NO 174<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium habana <400> SEQUENCE: 174 aacactccaa ttgggtgggg20 <210> SEQ ID NO 175 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium habana <400> SEQUENCE: 175ggttctcgtc tgtagtggac 20 <210> SEQ ID NO 176 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumhabana <400> SEQUENCE: 176 tgcacaacaa caggcaatcg 20 <210> SEQ ID NO 177<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium habana <400> SEQUENCE: 177 ggccgacttc ggttgaagtg20 <210> SEQ ID NO 178 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium habana <400> SEQUENCE: 178ggggtgtggt gtttgagtat 20 <210> SEQ ID NO 179 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumhabana <400> SEQUENCE: 179 gagcatctaa atgaacgcgt 20 <210> SEQ ID NO 180<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium habana <400> SEQUENCE: 180 tacgtgttcg ttttgtgtaa20 <210> SEQ ID NO 181 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium farcinogen <400> SEQUENCE:181 gttggtgatg gactgccaga 20 <210> SEQ ID NO 182 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumfarcinogen <400> SEQUENCE: 182 cgcgggaaag cgttggtgat 20 <210> SEQ ID NO183 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium farcinogen <400> SEQUENCE: 183 cgatgtcgtgggcagtagtg 20 <210> SEQ ID NO 184 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium farcinogen <400>SEQUENCE: 184 ggatatttca gccagcatct 20 <210> SEQ ID NO 185 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium farcinogen <400> SEQUENCE: 185 tgtctggtgg gtgcaaatga 20<210> SEQ ID NO 186 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium farcinogen <400> SEQUENCE:186 cggcgcggga aagcgttggt 20 <210> SEQ ID NO 187 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumfarcinogen <400> SEQUENCE: 187 gggaaagcgt tggtgatgga 20 <210> SEQ ID NO188 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium farcinogen <400> SEQUENCE: 188 tctagcacgcagagtgtggc 20 <210> SEQ ID NO 189 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium farcinogen <400>SEQUENCE: 189 tgggggcctt cgggtttctt 20 <210> SEQ ID NO 190 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium asiaticum <400> SEQUENCE: 190 catcccaatt ggtgggatgc 20<210> SEQ ID NO 191 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium asiaticum <400> SEQUENCE:191 cacaacaaca agcaagccag 20 <210> SEQ ID NO 192 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumasiaticum <400> SEQUENCE: 192 cactcgggcg ctagcacgaa 20 <210> SEQ ID NO193 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium asiaticum <400> SEQUENCE: 193 aactgatcgcgtcgccgttc 20 <210> SEQ ID NO 194 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium porcinum <400>SEQUENCE: 194 cagtagtgcg gttgggatat 20 <210> SEQ ID NO 195 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium porcinum <400> SEQUENCE: 195 ttcgattccc ccgccgtccg 20<210> SEQ ID NO 196 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium porcinum <400> SEQUENCE: 196cagccaggac ctgtagtggg 20 <210> SEQ ID NO 197 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumporcinum <400> SEQUENCE: 197 tggtgggtgc aaatgacaaa 20 <210> SEQ ID NO198 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium porcinum <400> SEQUENCE: 198 gagccggcgcgggaaagcgt 20 <210> SEQ ID NO 199 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium porcinum <400>SEQUENCE: 199 gtgatggaac tgctgaacac 20 <210> SEQ ID NO 200 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium porcinum <400> SEQUENCE: 200 ggctttgaga caacaggccc 20<210> SEQ ID NO 201 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium porcinum <400> SEQUENCE: 201cggggggtgg cattccgttg 20 <210> SEQ ID NO 202 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumporcinum <400> SEQUENCE: 202 cggcgtgttg ttgctccgct 20 <210> SEQ ID NO203 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium porcinum <400> SEQUENCE: 203 tagtggttgcgagcatctag 20 <210> SEQ ID NO 204 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium porcinum <400>SEQUENCE: 204 agtgtggctg ggggccttcg 20 <210> SEQ ID NO 205 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of probe or primer for detectingMycobacterium porcinum <400> SEQUENCE: 205 ggtcttgttg tgtgttgatg 20<210> SEQ ID NO 206 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: sequence ofprobe or primer for detecting Mycobacterium acapulcensis <400> SEQUENCE:206 tgccagcacc tgtagtgggt 20 <210> SEQ ID NO 207 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumacapulcensis <400> SEQUENCE: 207 gatcgtctgg tggggaaagc 20 <210> SEQ IDNO 208 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium acapulcensis <400> SEQUENCE: 208cgggcgtgaa agattgccag 20 <210> SEQ ID NO 209 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumacapulcensis <400> SEQUENCE: 209 agcccgtcgc ctctttgtcc 20 <210> SEQ IDNO 210 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium acapulcensis <400> SEQUENCE: 210cgagtgtggg atatcggaga 20 <210> SEQ ID NO 211 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumacapulcensis <400> SEQUENCE: 211 ggagcaccac gagacctgtt 20 <210> SEQ IDNO 212 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium acapulcensis <400> SEQUENCE: 212cgcccacatc gtgtgggagt 20 <210> SEQ ID NO 213 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumacapulcensis <400> SEQUENCE: 213 tcggtgactc aggcgattcg 20 <210> SEQ IDNO 214 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium acapulcensis <400> SEQUENCE: 214gggtgtggtg tttgatttgt 20 <210> SEQ ID NO 215 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumacapulcensis <400> SEQUENCE: 215 cgagcatctg aacgcagaga 20 <210> SEQ IDNO 216 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: sequence of probe orprimer for detecting Mycobacterium diernhoferi <400> SEQUENCE: 216acctgttgcc cgcccacatc 20 <210> SEQ ID NO 217 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: sequence of probe or primer for detecting Mycobacteriumdiernhoferi <400> SEQUENCE: 217 gtgtgggagt tcggtgactc 20 <210> SEQ ID NO218 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium diernhoferi <400> SEQUENCE: 218 ttcgttggatggcctcacac 20 <210> SEQ ID NO 219 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium diernhoferi<400> SEQUENCE: 219 tgcacaacaa actttgagaa 20 <210> SEQ ID NO 220 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium diernhoferi <400> SEQUENCE: 220 acaggccctgcggtgccgga 20 <210> SEQ ID NO 221 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium diernhoferi<400> SEQUENCE: 221 tcgttggagt cctggttgcc 20 <210> SEQ ID NO 222 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium diernhoferi <400> SEQUENCE: 222 ggccgcgagtcccggaagcg 20 <210> SEQ ID NO 223 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium diernhoferi<400> SEQUENCE: 223 gtctgttgtt gctccatctt 20 <210> SEQ ID NO 224 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium diernhoferi <400> SEQUENCE: 224 ggtgtggtgtttgatttgtg 20 <210> SEQ ID NO 225 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium diernhoferi<400> SEQUENCE: 225 atagtggttg cgagcatcta 20 <210> SEQ ID NO 226 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium diernhoferi <400> SEQUENCE: 226 acgcaagaggagtctgggtt 20 <210> SEQ ID NO 227 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium diernhoferi<400> SEQUENCE: 227 ttgttgtgtg tgtttgatgt 20 <210> SEQ ID NO 228 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium paratuberculosis <400> SEQUENCE: 228 gagcaccacgaaaagcaccc 20 <210> SEQ ID NO 229 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium paratuberculosis<400> SEQUENCE: 229 ctggtggggt gcgagccgtg 20 <210> SEQ ID NO 230 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium paratuberculosis <400> SEQUENCE: 230 gcgagccgtgaggggttccc 20 <210> SEQ ID NO 231 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium paratuberculosis<400> SEQUENCE: 231 aggggttccc gtctgtagtg 20 <210> SEQ ID NO 232 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium paratuberculosis <400> SEQUENCE: 232 acgggggccgggtgcgcaac 20 <210> SEQ ID NO 233 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium paratuberculosis<400> SEQUENCE: 233 caaatgattg ccagacacac 20 <210> SEQ ID NO 234 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium paratuberculosis <400> SEQUENCE: 234 tattgggcctgagacaacac 20 <210> SEQ ID NO 235 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium paratuberculosis<400> SEQUENCE: 235 cggtccgtcc gtgtggagtc 20 <210> SEQ ID NO 236 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium paratuberculosis <400> SEQUENCE: 236 tccatcttggtggtggggtg 20 <210> SEQ ID NO 237 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium paratuberculosis<400> SEQUENCE: 237 ggtgtttgag tattggatag 20 <210> SEQ ID NO 238 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium paratuberculosis <400> SEQUENCE: 238 tggttgcgagcatctagatg 20 <210> SEQ ID NO 239 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacterium paratuberculosis<400> SEQUENCE: 239 gcgcatggtc tccgtggccg 20 <210> SEQ ID NO 240 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of probe or primer fordetecting Mycobacterium paratuberculosis <400> SEQUENCE: 240 gttcatcgaaatgtgtaatt 20 <210> SEQ ID NO 241 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:sequence of probe or primer for detecting Mycobacteria sp. <400>SEQUENCE: 241 aaaagctgtt gtttgacggt 20 <210> SEQ ID NO 242 <211> LENGTH:25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: sequence of primer for amplifying MycobacteriaITS selected from the sequence of 16S rRNA in Mycobacteria <400>SEQUENCE: 242 tggatccgac gaagtcgtaa caagg 25 <210> SEQ ID NO 243 <211>LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: sequence of primer for amplifyingMycobacteria ITS selected from the sequence of 23S rRNA in Mycobacteria<400> SEQUENCE: 243 tggatcctgc caaggcatcc accat 25

What is claimed is:
 1. An isolated DNA molecule of an Internal Transcribed Spacer region of Mycobacterium fortuitum where said DNA molecule consists of, SEQ ID NO:
 1. 2. An oligonucleotide for detection and identification of Mycobacterium fortuitum wherein said oligonucltide is selected from the group consisting of SEQ ID NOs: 28 to
 38. 3. A method for detecting Mycobacterium fortuitum from a sample comprising: obtaining a sample DNA from the sample; hybridizing the sample DNA with a DNA primer, the DNA primer is at least one oligonucleotide selected from the group consisting of SEQ ID Nos: 28 to 38, amplifying the sample DNA hybridized with the DNA primer; and detecting and identifying a product of amplification thereby detecting and identifying Mycobacterium fortuitum. 